Wieler L H, Szattelberger C, Weiss R, Bauerfeind R, Kutzer P, Failing K, Baljer G
Institut für Hygiene und Infektionskrankheiten der Tiere, Justus-Liebig-Universität Giessen, Germany.
Berl Munch Tierarztl Wochenschr. 1999 Dec;112(12):465-71.
Dog sera (n = 118) were tested for antibodies recognizing Borrelia (B.) burgdorferi sensu stricto strain B31 (ATCC 35210) antigens. In total, 18 of the dog sera gave positive results in a whole cell sonicate ELISA (WCS ELISA). These positive sera were further evaluated by immunoblot assay, utilizing a whole bacterial lysate as antigens. 94.4% (17 of 18) of the dog sera reacted with immunodominant antigens at 20-22 kDa (protein C, pC), 31 kDa (outer surface protein A, OspA), 34 kDa (outer surface protein B, OspB), 41 kDa (flagellin), 60 kDa ("common antigen"), and/or 100 kDa (presumably p100). Sera recognizing pC (20-22 kDa) and antigens > 94 kDa always detected the highest number of antigen bands, indicating the specificity of those antigens in serological diagnosis. The results clearly demonstrate that the WCS ELISA is a useful tool for testing sera of dogs for antibodies against B. burgdorferi. However, positive results should be confirmed by immunoblot, using WCS as antigen. According to the presented data, we recommend criteria for B. burgdorferi immunoblots using dog sera as follows: sera have to be considered as positive if they detect the 41 kDa flagellin, and two of the 5 immunodominant antigens, namely > 94 kDa (presumably p100), 60 kDa ("common antigen"), 34 kDa and 29-31 kDa (OspB and OspA, respectively) and 20-22 kDa (pC). If sera only recognize the 41 kDa flagellin, this result is equivocal, requiring testing a second serum sample 4 to 8 weeks later.
检测了118份犬血清中识别伯氏疏螺旋体狭义种B31菌株(ATCC 35210)抗原的抗体。总共18份犬血清在全细胞超声裂解物酶联免疫吸附测定(WCS ELISA)中呈阳性结果。利用全细菌裂解物作为抗原,通过免疫印迹分析对这些阳性血清进行了进一步评估。94.4%(18份中的17份)的犬血清与20 - 22 kDa(蛋白C,pC)、31 kDa(外膜蛋白A,OspA)、34 kDa(外膜蛋白B,OspB)、41 kDa(鞭毛蛋白)、60 kDa(“共同抗原”)和/或100 kDa(可能是p100)的免疫显性抗原发生反应。识别pC(20 - 22 kDa)和大于94 kDa抗原的血清总是检测到最多数量的抗原条带,表明这些抗原在血清学诊断中的特异性。结果清楚地表明,WCS ELISA是检测犬血清中抗伯氏疏螺旋体抗体的有用工具。然而,阳性结果应以WCS作为抗原通过免疫印迹进行确认。根据所呈现的数据,我们推荐使用犬血清进行伯氏疏螺旋体免疫印迹的标准如下:如果血清检测到41 kDa鞭毛蛋白以及5种免疫显性抗原中的两种,即大于94 kDa(可能是p100)、60 kDa(“共同抗原”)、34 kDa和29 - 31 kDa(分别为OspB和OspA)以及20 - 22 kDa(pC),则该血清应被视为阳性。如果血清仅识别41 kDa鞭毛蛋白,则该结果不明确,需要在4至8周后检测第二份血清样本。